Epothilone A (Rxe2x95x90H) and Epothilone B (Rxe2x95x90CH3) are produced by Sorangium cellulosum strain So ce 90, the structures of which are shown below, and were the first of several epothilones to be isolated and characterized. Hofle et al., 1996, Angew. Chem. Int. Ed. Engl. 35(13/14): 1567-1569. 
Epothilone A and epothilone B possess many of the advantageous properties of taxol. As a result, there is significant interest in these and structurally related compounds as potential chemotherapeutic agents. The desoxy counterparts of epothilones A and B are known as epothilone C (Rxe2x95x90H) and epothilone D (Rxe2x95x90CH3), and also exhibit similar anti-tumor activity but with less cytotoxicity. The structures of epothilones C and D are shown below. 
Although other naturally occurring epothilones have been described in the literature, these compounds are produced in exceedingly small amounts. For example, PCT publication WO 99/65913 describes 39 naturally occurring epothilones obtained from Sorangium cellulosum So ce 90 of which epothilones A, B, C, and D together account for approximately 98.9% of the total epothilones produced. The 35 other naturally occurring epothilone compounds together account for the remaining 1.1% and include epothilone C6 (which may also be referred to as 10,11-dehydroepothilone C) and whose structure is shown below 
Due to the increasing interest in epothilones as anti-cancer agents, novel derivatives of these compounds are needed and desired to more fully develop their therapeutic potential.
The present invention relates to 16-membered macrocyclic compounds. In one aspect of the present invention, 16-membered macrocyclic compounds having a conjugated diene are provided. In another aspect of the present invention, compounds of the following formula 
are provided wherein:
R1, R2, R3, and R5 are each independently hydrogen, C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, aryl or alkylaryl;
R4 is hydrogen, C1-C10 alkyl, C1-C10 hydroxyalkyl, C1-C10 haloalkyl, aryl, xe2x80x94C(xe2x95x90O)R6, xe2x80x94C(xe2x95x90O)OR6, xe2x80x94NR6R7 where R6 and R7 are each independently hydrogen, C1-C10 aliphatic, aryl or alkylaryl;
W is O, NR8 where R8 is hydrogen, C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, aryl or alkylaryl;
X is O, CH2 or a carbon-carbon double bond;
Y is absent or a C1-C10 alkyl, C2-C10 alkenyl, or C2-C10 alkynyl; and
Ar is aryl; provided that 10,11-dehydroepothilone C is excluded. In another aspect of the present invention, methods for using the inventive compounds are provided.
The present invention relates to novel compounds that are useful for the treatment of cancer and other conditions characterized by abnormal cellular proliferation in a subject in need thereof.
Definitions
Statements regarding the scope of the present invention and definitions of terms used herein are listed below. The definitions apply to the terms as they are used throughout this specification, unless otherwise limited in specific instances, either individually or as part of a larger group.
All stereoisomers of the inventive compounds are included within the scope of the invention, as pure compounds as well as mixtures thereof. Individual enantiomers, diastereomers, geometric isomers, and combinations and mixtures thereof are all encompassed by the present invention. Furthermore, some of the crystalline forms for the compounds may exist as polymorphs and as such are included in the present invention. In addition, some of the compounds may form solvates with water (i.e., hydrates) or common organic solvents, and such solvates are also encompassed within the scope of this invention.
Protected forms of the inventive compounds are included within the scope of the present invention. A variety of protecting groups are disclosed, for example, in T. H. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, Third Edition, John Wiley and Sons, New York (1999), which is incorporated herein by reference in its entirety. For example, a hydroxy protected form of the inventive compounds are those where at least one of the hydroxyl groups is protected by a hydroxy protecting group. Illustrative hydroxyl protecting groups include but not limited to tetrahydropyranyl; benzyl; methylthiomethyl; ethylthiomethyl; pivaloyl; phenylsulfonyl; triphenylmethyl; trisubstituted silyl such as trimethyl silyl, triethylsilyl, tributylsilyl, tri-isopropylsilyl, t-butyldimethylsilyl, tri-t-butylsilyl, methyldiphenylsilyl, ethyldiphenylsilyl, t-butyldiphenylsilyl and the like; acyl and aroyl such as acetyl, pivaloylbenzoyl, 4-methoxybenzoyl, 4-nitrobenzoyl and aliphatic acylaryl and the like. Keto groups in the inventive compounds may similarly be protected.
The present invention includes within its scope prodrugs of the compounds of this invention. In general, such prodrugs are functional derivatives of the compounds that are readily convertible in vivo into the required compound. Thus, in the methods of treatment of the present invention, the term xe2x80x9cadministeringxe2x80x9d shall encompass the treatment of the various disorders described with the compound specifically disclosed or with a compound which may not be specifically disclosed, but which converts to the specified compound in vivo after administration to a subject in need thereof. Conventional procedures for the selection and preparation of suitable prodrug derivatives are described, for example, in xe2x80x9cDesign of Prodrugsxe2x80x9d, H. Bundgaard ed., Elsevier, 1985.
As used herein, the term xe2x80x9caliphaticxe2x80x9d refers to saturated and unsaturated straight chained, branched chain, cyclic, or polycyclic hydrocarbons that may be optionally substituted at one or more positions. Illustrative examples of aliphatic groups include alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, and cycloalkynyl moieties. The term xe2x80x9calkylxe2x80x9d refers to straight or branched chain saturated hydrocarbon substituent. xe2x80x9cAlkenylxe2x80x9d refers to a straight or branched chain hydrocarbon substituent with at least one carbon-carbon double bond. xe2x80x9cAlkynylxe2x80x9d refers to a straight or branched chain hydrocarbon substituent with at least one carbon-carbon triple bound.
The term xe2x80x9carylxe2x80x9d refers to monocyclic or polycyclic groups having at least one aromatic ring structure that optionally include one or more heteroatoms and preferably include three to fourteen carbon atoms. Aryl substituents may optionally be substituted at one or more positions. Illustrative examples of aryl groups include but are not limited to: furanyl, imidazolyl, indanyl, indenyl, indolyl, isooxazolyl, isoquinolinyl, naphthyl, oxazolyl, oxadiazolyl, phenyl, pyrazinyl, pyridyl, pyrimidinyl, pyrrolyl, pyrazolyl, quinolyl, quinoxalyl, tetrahydronaphththyl, tetrazolyl, thiazolyl, thienyl, and the like.
The aliphatic (i.e., alkyl, alkenyl, etc.) and aryl moieties may be optionally substituted with one or more substituents, preferably from one to five substituents, more preferably from one to three substituents, and most preferably from one to two substituents. The definition of any substituent or variable at a particular location in a molecule is independent of its definitions elsewhere in that molecule. It is understood that substituents and substitution patterns on the compounds of this invention can be selected by one of ordinary skill in the art to provide compounds that are chemically stable and that can be readily synthesized by techniques known in the art as well as those methods set forth herein. Examples of suitable substituents include but are not limited to: alkyl, alkenyl, alkynyl, aryl, halo; trifluoromethyl; trifluoromethoxy; hydroxy; alkoxy; cycloalkoxy; heterocyclooxy; oxo; alkanoyl (xe2x80x94C(xe2x95x90O)-alkyl which is also referred to as xe2x80x9cacylxe2x80x9d)); aryloxy; alkanoyloxy; amino; alkylamino; arylamino; aralkylamino; cycloalkylamino; heterocycloamino; disubstituted amines in which the two amino substituents are selected from alkyl, aryl, or aralkyl; alkanoylamino; aroylamino; aralkanoylamino; substituted alkanoylamino; substituted arylamino; substituted aralkanoylamino; thiol; alkylthio; arylthio; aralkylthio; cycloalkylthio; heterocyclothio; alkylthiono; arylthiono; aralkylthiono; alkylsulfonyl; arylsulfonyl; aralkylsulfonyl; sulfonamido (e.g., SO2NH2); substituted sulfonamido; nitro; cyano; carboxy; carbamyl (e.g., CONH2); substituted carbamyl (e.g., xe2x80x94C(xe2x95x90O)NRRxe2x80x2 where R and Rxe2x80x2 are each independently hydrogen, alkyl, aryl, aralkyl and the like); alkoxycarbonyl, aryl, substituted aryl, guanidino, and heterocyclo such as indoyl, imidazolyl, furyl, thienyl, thiazolyl, pyrrolidyl, pyridyl, pyrimidyl and the like. Where applicable, the substituent may be further substituted such as with, alkyl, alkoxy, aryl, aralkyl, halogen, hydroxy and the like.
The terms xe2x80x9calkylarylxe2x80x9d or xe2x80x9carylalkylxe2x80x9d refer to an aryl group with an aliphatic substituent that is bonded to the compound through the aliphatic group. An illustrative example of an alkylaryl or arylalkyl group is benzyl, a phenyl with a methyl group that is bonded to the compound through the methyl group (xe2x80x94CH2Ph where Ph is phenyl).
The term xe2x80x9cacylxe2x80x9d refers to xe2x80x94C(xe2x95x90O)R where R is an aliphatic group, preferably a C1-C6 moiety.
The term xe2x80x9calkoxyxe2x80x9d refers to xe2x80x94OR wherein O is oxygen and R is an aliphatic group.
The term xe2x80x9caminoalkylxe2x80x9d refers to xe2x80x94RNH2 where R is an aliphatic moiety.
The terms xe2x80x9chalogen,xe2x80x9d xe2x80x9chaloxe2x80x9d, or xe2x80x9chalidexe2x80x9d refer to fluorine, chlorine, bromine and iodine.
The term xe2x80x9chaloalkylxe2x80x9d refers to xe2x80x94RX where R is an aliphatic moiety and X is one or more halogens.
The term xe2x80x9chydroxyalkylxe2x80x9d refers to xe2x80x94ROH where R is an aliphatic moiety.
The term xe2x80x9coxoxe2x80x9d refers to a carbonyl oxygen (xe2x95x90O).
In addition to the explicit substitutions at the above-described groups, the inventive compounds may include other substitutions where applicable. For example, the lactone or lactam backbone or backbone substituents may be additionally substituted (e.g., by replacing one of the hydrogens or by derivatizing a non-hydrogen group) with one or more substituents such as C1-C5 aliphatic, C1-C5 alkoxy, aryl, or a functional group. Illustrative examples of suitable functional groups include but are not limited to: acetal, alcohol, aldehyde, amide, amine, boronate, carbamate, carboalkoxy, carbonate, a carbodiimide, carboxylic acid, cyanohydrin, disulfide, enamine, ester, ether, halogen, hydrazide, hydrazone, imide, imido, imine, isocyanate, ketal, ketone, nitro, oxime, phosphine, phosphonate, phosphonic acid, quaternary ammonium, sulfenyl, sulfide, sulfone, sulfonic acid, thiol, and the like.
The term xe2x80x9cpurifiedxe2x80x9d as used herein to refer to a compound of the present invention, means that the compound is in a preparation in which the compound forms a major component of the composition, such as constituting about 50%, about 60%, about 70%, about 80%, about 90%, about 95% or more by weight of the components in the composition.
The term xe2x80x9csubjectxe2x80x9d as used herein, refers to an animal, preferably a mammal, who has been the object of treatment, observation or experiment and most preferably a human who has been the object of treatment and/or observation.
The term xe2x80x9ctherapeutically effective amountxe2x80x9d as used herein, means that amount of active compound or pharmaceutical agent that elicits the biological or medicinal response in a tissue system, animal or human that is being sought by a researcher, veterinarian, medical doctor or other clinician, which includes alleviation of the symptoms of the disease or disorder being treated.
The term xe2x80x9ccompositionxe2x80x9d is intended to encompass a product comprising the specified ingredients in the specified amounts, as well as any product that results, directly or indirectly, from combinations of the specified ingredients in the specified amounts.
The term xe2x80x9cpharmaceutically acceptable saltxe2x80x9d is a salt of one or more of the inventive compounds. Suitable pharmaceutically acceptable salts of the compounds include acid addition salts which may, for example, be formed by mixing a solution of the compound with a solution of a pharmaceutically acceptable acid such as hydrochloric acid, sulfuric acid, fumaric acid, maleic acid, succinic acid, acetic acid, benzoic acid, citric acid, tartaric acid, carbonic acid or phosphoric acid. Furthermore, where the compounds of the invention carry an acidic moiety, suitable pharmaceutically acceptable salts thereof may include alkali metal salts (e.g., sodium or potassium salts); alkaline earth metal salts (e.g., calcium or magnesium salts); and salts formed with suitable organic ligands (e.g., ammonium, quaternary ammonium and amine cations formed using counteranions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, alkyl sulfonate and aryl sulfonate). Illustrative examples of pharmaceutically acceptable salts include but are not limited to:
acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, butyrate, calcium edetate, camphorate, camphorsulfonate, camsylate, carbonate, chloride, citrate, clavulanate, cyclopentanepropionate, digluconate, dihydrochloride, dodecylsulfate, edetate, edisylate, estolate, esylate, ethanesulfonate, formate, fumarate, gluceptate, glucoheptonate, gluconate, glutamate, glycerophosphate, glycolylarsanilate, hemisulfate, heptanoate, hexanoate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroiodide, 2-hydroxy-ethanesulfonate, hydroxynaphthoate, iodide, isothionate, lactate, lactobionate, laurate, lauryl sulfate, malate, maleate, malonate, mandelate, mesylate, methanesulfonate, methylsulfate, mucate, 2-naphthalenesulfonate, napsylate, nicotinate, nitrate, N-methylglucamine ammonium salt, oleate, oxalate, pamoate (embonate), palmitate, pantothenate, pectinate, persulfate, 3-phenylpropionate, phosphate/diphosphate, picrate, pivalate, polygalacturonate, propionate, salicylate, stearate, sulfate, subacetate, succinate, tannate, tartrate, teoclate, tosylate, triethiodide, undecanoate, valerate, and the like.
The term xe2x80x9cpharmaceutically acceptable carrierxe2x80x9d is a medium that is used to prepare a desired dosage form of the inventive compound. A pharmaceutically acceptable carrier includes solvents, diluents, or other liquid vehicle; dispersion or suspension aids; surface active agents; isotonic agents; thickening or emulsifying agents, preservatives; solid binders; lubricants and the like. Remington""s Pharmaceutical Sciences, Fifteenth Edition, E. W. Martin (Mack Publishing Co., Easton, Pa., 1975) and Handbook of Pharmaceutical Excipients, Third Edition, A. H. Kibbe, ed. (Amer. Pharmaceutical Assoc. 2000), both of which are incorporated herein by reference in their entireties, disclose various carriers used in formulating pharmaceutical compositions and known techniques for the preparation thereof.
The term xe2x80x9cpharmaceutically acceptable esterxe2x80x9d is an ester that hydrolzyes in vivo into a compound of the present invention or a salt thereof. Illustrative examples of suitable ester groups include, for example, those derived from pharmaceutically acceptable aliphatic carboxylic acids such as formates, acetates, propionates, butyrates, acrylates, and ethylsuccinates.
Compounds of the Present Invention
In one aspect of the present invention, the compounds are a 16 membered macrocycle having a conjugated diene. The macrocycle may be a cyclic ester or lactone, a cyclic amide or lactam, or a cyclic thioester.
In another aspect of the present invention, compounds of the following formula 
are provided wherein:
R1, R2, R3, and R5 are each independently hydrogen, C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, aryl or alkylaryl;
R4 is hydrogen, halogen, C1-C10 alkyl, C1-C10 hydroxyalkyl, C1-C10 haloalkyl, aryl, xe2x80x94C(xe2x95x90O)R6, xe2x80x94C(xe2x95x90O)OR6, xe2x80x94NR6R7 where R6 and R7 are each independently hydrogen, C1-C10 aliphatic, aryl or alkylaryl;
W is O, NR8 where R8 is hydrogen, C1-C10 alkyl, C2-C10 alkenyl, C2-C10 alkynyl, aryl or alkylaryl;
X is O, CH2 or a carbon-carbon double bond;
Y is absent or a C1-C10 alkyl, C2-C10 alkenyl, or C2-C10 alkynyl; and
Ar is aryl; provided that 10,11-dehydroepothilone C is excluded.
In one embodiment, compounds of formula I are provided wherein:
R1, R2, R3, and R5 are each independently hydrogen or C1-C5 alkyl;
R4 is hydrogen, C1-C5 alkyl, C1-C5 hydroxyalkyl, C1-C5 haloalkyl, xe2x80x94C(xe2x95x90O)R6, xe2x80x94C(xe2x95x90O)OR6, xe2x80x94NR6R7 where R6 and R7 are each independently hydrogen or C1-C5 alkyl;
W is O or NR8 where R8 is hydrogen or C1-C5 alkyl;
X is O, CH2 or a carbon-carbon double bond;
Y is absent or C2-C5 alkenyl; and
Ar is a heteroaryl.
In another embodiment, compound of formula I are provided wherein:
R1, R2, R3, and R5 are each independently hydrogen, methyl or ethyl;
R4 is hydrogen, methyl, ethyl, hydroxymethyl, hydroxyethyl, fluoromethyl or fluoroethyl;
W is O or NH;
X is O, CH2 or a carbon-carbon double bond;
Y is absent or C2-C5 alkenyl; and
Ar is an aryl selected from the group consisting of phenyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, furyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, thiazolyl, oxazolyl, quinolyl, isoquinolyl, quinoxalyl, indolyl, benzothiazolyl, benzoxazolyl, benzoimidazolyl, and benzopyrazolyl.
In another embodiment, compound of formula I are provided wherein:
R1, R2, R3, and R5 are each independently hydrogen or methyl;
R4 is hydrogen, methyl, ethyl, or fluoromethyl;
W is O or NH;
X is O, CH2 or a carbon-carbon double bond;
Y is absent or C2-C3 alkenyl; and
Ar is an aryl selected from the group consisting of thiazolyl, oxazolyl, quinolyl, isoquinolyl, quinoxalyl, indolyl, benzothiazolyl, benzoxazolyl, benzoimidazolyl, and benzopyrazolyl.
In another aspect of the present invention, compounds of the following formula 
are provided wherein:
W is O or NR8 where R8 is hydrogen or C1-C5 alkyl;
X is O, CH2 or a carbon-carbon double bond;
R4 is hydrogen, C1-C5 alkyl, C1-C5 hydroxyalkyl, C1-C5 haloalkyl, xe2x80x94C(xe2x95x90O)R6, xe2x80x94C(xe2x95x90O)OR6, xe2x80x94NR6R7 where R6 and R7 are each independently hydrogen or C1-C5 alkyl; and,
R9 is aryl or R11CHxe2x95x90C(R10)xe2x80x94 wherein R11 is aryl and R10 is hydrogen, halide, C1-C5 alkyl, C1-C5 hydroxylalkyl, or C1-C5 haloalkyl.
In another embodiment, compounds of formula II are provided wherein:
W is O or NH; and,
X is CH2 or a carbon-carbon double bond;
R4 is hydrogen, methyl, ethyl, hydroxymethyl, hydroxyethyl, fluoromethyl or fluoroethyl; and,
R9 is a bicyclic heteroaryl or R11CHxe2x95x90C(R10)xe2x80x94 wherein R11 is 2-methyl-1,3-thiazolinyl, 2-methyl-13, oxazolinyl, 2-hydroxymethyl-1,3-thiazolinyl, or 2-hydroxymethyl-1,3-oxazolinyl and R10 is hydrogen or methyl.
In another embodiment, compounds of formula II are provided wherein:
W is O or NH; and,
X is CH2 or a carbon-carbon double bond;
R4 is hydrogen, methyl, ethyl, hydroxymethyl, hydroxyethyl, fluoromethyl or fluoroethyl; and,
R9 is selected from the group consisting of 
In another aspect of the present invention, compounds of the following formulas 
are provided wherein:
R1, R2, R3, and R5 are each independently hydrogen or C1-C5 alkyl; and,
R4 is hydrogen, C1-C5 alkyl, C1-C5 hydroxyalkyl, C1-C5 haloalkyl, xe2x80x94C(xe2x95x90O)R6, xe2x80x94C(xe2x95x90O)OR6, xe2x80x94NR6R7 where R6 and R7 are each independently hydrogen or C1-C5 alkyl;
R12 is hydrogen, halide, C1-C5 alkyl, C1-C5 hydroxyalkyl, C1-C5 haloalkyl, xe2x80x94(CH2)mC(xe2x95x90O)R6, xe2x80x94(CH2)mC(xe2x95x90O)OR6, xe2x80x94(CH2)mNR6R7 where m is 0, 1 or 2, and R6 and R7 are each independently hydrogen or C1-C5 alkyl; provided that 10,11-dehydroepothilone C is excluded.
In one embodiment, compounds are of formulas III or IV wherein
R1, R2, R3, and R5 are each independently hydrogen, methyl or ethyl;
R4 is methyl, ethyl, hydroxymethyl, fluoromethyl, hydroxyethyl, or fluoroethyl; and,
R12 is hydrogen, C1-C3 alkyl, C1-C3 hydroxyalkyl, C1-C3 haloalkyl, xe2x80x94(CH2)mC(xe2x95x90O)R6, xe2x80x94(CH2)mC(xe2x95x90O)OR6, xe2x80x94(CH2)mNR6R7 where m is 0 or 1, and R6 and R7 are each independently hydrogen or methyl.
In another embodiment, compounds are of formulas III or IV wherein
R1, R3, and R5 are each hydrogen;
R2 is methyl;
R4 is methyl, ethyl, or fluoromethyl; and,
R12 is methyl, hydroxymethyl, fluoromethyl, and aminomethyl.
In another aspect of the present invention, the following compounds are provided: 
The compounds of the present invention are cytotoxic agents and may be used in any suitable manner including but not limited to as anti-cancer agents. An illustrative assay for assessing the degree of cytotoxicy and tubulin polyermization is described in Example 1.
Synthetic Methods
General principles of organic chemistry including functional moieties and reactivity and common protocols are described by for example in Advanced Organic Chemistry 3rd Ed. by Jerry March (1985) which is incorporated herein by reference in its entirety. In addition, it will be appreciated by one of ordinary skill in the art that the synthetic methods described herein may use a variety of protecting groups whether or not they are explicitly described. A xe2x80x9cprotecting groupxe2x80x9d as used herein means a moiety used to block functional moiety such as oxygen, sulfur, or nitrogen so that a reaction can be carried out selectively at another reactive site in a multifunctional compound. General principles including specific functional groups and their uses are described for example in T. H. Greene and P. G. M. Wuts, Protective Groups in Organic Synthesis, 3rd edition, John Wiley and Sons, New York (1999).
In one aspect of the present invention, two fragments are coupled together and subsequently cyclized to form 16 membered macrocycles of the present invention. In one embodiment, fragments A and B are joined together under Heck coupling conditions (a palladium catalyst such as (diphenylphosphineferrocenyl)dichloropalladium; a base such as cesium carbonate or 9-borabicyclo[3.3.1]nonane; and triphenylarsine). In another embodiment, fragments A and Bxe2x80x2 (an alkyne form of fragment B) are joined together under Suzuki coupling reaction conditions. Fragment Bxe2x80x2 is treated with a borane such as catechol borane or 9-borabicyclo[3.3.1]nonane, and then fragments Bxe2x80x2 and A, a palladium catalyst such as (diphenylphosphineferrocenyl)dichloropalladium, a base such as cesium carbonate, and triphenylarsine are reacted together. Both methods are illustrated schematically by Scheme 1 where Ar, W, Y, R1, R2, R3, R4, R5 are as described previously and Z is a protected hydroxy group or a protected amino group that is capable of becoming W upon deprotection and cyclization. 
Fragment A is made using a number of methods. Scheme 2A illustrates one embodiment where R5 is hydrogen and Z is a protected hydroxy group and Arxe2x80x94Y, and R4 are as described previously. 
Briefly, (2R)-N-acetyl-2,10-camphorsultam is treated with a dialkylboron triflate such as diethylborontriflate and a base such as diisopropyl ethylamine and then reacted with Arxe2x80x94Yxe2x80x94CHO in an Oppolzer aldol condensation. The resulting alcohol is protected by reacting the compound with triethylsilyl triflate and lutidine and reduced to form the aldehyde. Fragment A is formed by extending the aldehyde in a Wittig reaction by treating the aldehyde with a phosphonium salt such as iodoethyltriphenylphosphonium iodide prepared in situ from ethyltriphenylphosphonium iodide.
Scheme 2B illustrates another embodiment where Arxe2x80x94Y, R4 and R5 are as described previously. This method is preferred where R5 is a non-hydrogen moiety. 
Aldehyde Arxe2x80x94Yxe2x80x94CHO is treated with diisopinocampheyl-allylborane in a Brown asymmetric allylation. The resulting alcohol is protected with triethylsilyl triflate and lutidine and the alkene is oxidized to an aldehyde. The aldehyde is extended as in Scheme 2A in a Wittig reaction with a phosphonium salt such as iodoethyltriphenylphosphonium iodide prepared in situ from ethyltriphenylphosphonium iodide.
Scheme 3 illustrates another embodiment where Z is a protected amino group and Arxe2x80x94Y, R4 and R5 are as described previously. 
Briefly, the hydroxy protected form of fragment A of the opposite configuration as that described in Schemes 2A and 2B is deprotected and treated with diphenylphosphoryl azide and diazabicycloundecene to form the azido-version of fragment A.
Fragment B where R1 is hydrogen is prepared as described by Scheme 4A. 
A 1,1-diisopropoxy-2,2-dimethyl-3-alkanone is extended in an aldol condensation reaction. The resulting hydroxyl group is protected with trichloroethoxycarbonyl chloride, and the acetal protecting group is removed to yield a C3-C11 Intermediate of fragment B. This intermediate aldehyde is extended in another aldol reaction and protected to yield fragment B.
Fragment Bxe2x80x2 (where R3 is hydrogen) and where R1 is hydrogen is prepared as described by Scheme 4B. 
The method is similar to that described by Scheme 4A except that the first aldol reaction is performed using the alkynal 
instead of the alkenal 
Fragments B and Bxe2x80x2 where R1 is a non-hydrogen is prepared as described by Scheme 5. 
The alkene and alkyne versions of the C3-C11 Intermediate are prepared as described by Schemes 4A and 4B respectively. The C3-C11 Intermediate is treated with N-propionyl-benzyloxazolidinone in an Evans aldol reaction. The resulting alcohol is protected with the silyl group and then benzyloxazlidinone is hydrolyzed. Esterification with with tert-butanol yields fragment B.
Fragments A and B can be joined together in a Heck coupling reaction to form a diene as shown by Scheme 6A. 
Alternatively, Fragments A and Bxe2x80x2 can be joined together in a Suzuki coupling reaction to form the same diene as shown by Scheme 6B. 
Compounds of the present invention that are cyclic lactones are made from the coupling of fragment A (where Z is a protected hydroxyl group) and fragment B (or Bxe2x80x2) as illustrated by Scheme 7A. 
The diene product of either the Heck coupling of fragments A and B or the Suzuki coupling of fragments A and Bxe2x80x2 is subjected to an ester exchange and then partially deprotected. The resulting product is lactonized and deprotected to yield compounds corresponding to formula I where W is O.
Compounds of the present invention that are cyclic lactams are made from the coupling of fragment A (where Z is a protected amino group such as N3) and fragment B (or Bxe2x80x2) as illustrated by Schemes 7B and 7C. 
The diene product of either the Heck coupling of fragments A and B or the Suzuki coupling of fragments A and Bxe2x80x2 is subjected to a Staudinger reduction and the resulting amine is protected. The resulting product is treated with trifluoroacetic acid and then cyclized to form the cyclic lactam. Deprotection of the Troc protecting group yields compounds corresponding to formula I where W is NH.
Methods for making N-alkyl lactam derivatives are described by Scheme 7C. 
The amino-carboxyacid is made as described by Scheme 7B and then subjected to reductive amination. The resulting product is then cyclized and deprotected to yield compounds corresponding to formula I where W is NR8.
In another aspect of the present invention, the inventive compounds are made from modified versions of fragments A and B (designated as Axe2x80x3 and Bxe2x80x3) using Stille coupling. 
As shown by Scheme 8, the Stille coupling reaction of fragments Axe2x80x3 and Bxe2x80x3 makes the same intermediate as that produced from the previously described Heck coupling or Suzuki coupling reactions. This intermediate can be cyclized under conditions similar to that described by Schemes 7A, 7B, and 7C to make the compounds of the present invention.
Fragment Axe2x80x3 can be made as described by Scheme 9 by stannylation of fragment A. 
Fragment Bxe2x80x3 where R3 is hydrogen is made by can be made starting with fragment Bxe2x80x2. 
As shown by Scheme 10A, fragment Bxe2x80x2 is treated with catechol borane in a hydroboration reaction and then iodinated to yield fragment Bxe2x80x3 where R3 is hydrogen.
Fragment Bxe2x80x3 where R3 is a non-hydrogen is also made starting from fragment Bxe2x80x2 
As shown by Scheme 10B, fragment Bxe2x80x2 is treated wtih zirconocene dichloride and trialkylaluminum in a Schwartz reaction to yield fragment Bxe2x80x3 where R3 is alkyl.
Biological Methods
In another aspect of the present invention, a subset of the inventive compounds is made using biological methods. In one embodiment, 10,11-dehydroepothilone D, whose structure is shown below, 
is isolated from a strain of Myxococcus xanthus, K111-40-1. This strain expresses the epothilone polyketide synthase but not an active epoK gene product and so produces primarily epothilone D and lesser amounts of epothilone C and 10,11-dehydroepothilone D. Strain K111-40-1 (PTA-2712) was deposited with the American Type Culture Collection (xe2x80x9cATCCxe2x80x9d), 10801 University Blvd., Manassas, Va., 20110-2209 USA, on Nov. 21, 2000. In another embodiment, 10,11-dehydroepothilone D may be isolated from M. xanthus strain K111-72-4.4 that expresses the epothilone polyketide synthase and contains an epoK gene with an inactivating in frame deletion. Strain K111-72-4.4 (PTA-2713) also was deposited with the ATCC on Nov. 21, 2000. Methods for fermentation of these strains, purification of 10,11-dehydroepothilone D produced by these strains, and recombinant strains that make 10,11-dehydroepothilone D are described in related application U.S. Ser. No. 09/825,876 filed on Apr. 3, 2001, by inventors Robert Arslanian, John Carney and Brian Metcalf entitled EPOTHILONE COMPOUNDS AND METHODS FOR MAKING AND USING THE SAME.
Recombinant techniques can be used to make subset of the compounds of the present invention. These compounds include those with substituents at the C-2 and/or C-6 and/or C-8 and/or C-10 and/or C-14 positions that differ from the naturally occurring epothilones A-D. Procedures for making these kinds of changes in heterologous hosts such as Myxococcus xanthus, Steptomyces lividans, and Pseudomonas fluorescens are described in U.S. Pat. No. 6,303,342, entitled RECOMBINANT METHODS AND MATERIALS FOR PRODUCING EPOTHILONE AND EPOTHILONE DERIVATIVES, which is incorporated herein by reference. Among other things, the patent provides the nucleotide sequence of the epothilone PKS and modification enzyme genes cloned from Sorangium cellulosum SMP44; cosmids containing overlapping fragments of the epothilone PKS and modification enzyme genes; plasmids having the full complement of epoA, epoB, epoC, epoD, epoE, epoF, epoK, and epoL genes; and heterologous host cells for making epothilones and epothilone derivatives. Cosmids, pKOS35-70.1A2 (ATCC 203782), pKOS35-70.4 (ATCC 203781), pKOS35-70.8A3 (ATCC 203783), and pKOS35-79.85 (ATCC 203780); plasmid pair, pKOS039-124R (PTA-926) and pKOS039-126R (PTA-927); and strain K111-32.25 (PTA-1700) derived from Myxococcus xanthus containing all the epothilone genes and their promoters, have been deposited with the ATCC on April 14, 2000. Additional procedures for making epothilones in Myxococcus are described in: U.S. Ser. No. 09/560,367 filed Apr. 28, 2000; No. 60/232,696, filed Sep. 14, 2000, now lapsed; No. 60/257,517 filed Dec. 21, 2000, now lapsed; and No. 60/269,020, filed Feb. 13, 2001, all of which are entitled PRODUCTION OF POLYKETIDES and are also incorporated herein by reference. Illustrative examples of compounds that may be made using recombinant techniques include: 2-methyl-10,11-dehydroepothilone C or D; 6-desmethyl-10,11-dehydroepothilone C or D; 8-desmethyl-10,11-dehydroepothilone C or D; 10-methyl-10,11-dehydroepothilone C or D; and 14-methyl-10,11-dehydroepothilone C or D.
In another aspect of the present invention, biologically derived strategies are used to modify certain compounds of the present invention regardless of whether the compounds are made biologically or by de novo chemical synthesis. In one embodiment, a microbially-derived hydroxylase is used to hydroxylate a terminal alkane, particularly an alkyl substituent of the thiazole moiety of the inventive compounds. Protocols for effectuating such a transformation are described for example by PCT Publication No. WO 00/39276 which is incorporated herein in its entirety by reference. Example 26 describes in greater detail the hydroxylation of the C-20 methyl of 10,11-dehydroepothilone D to 21-hydroxy-10,11-dehydroepothilone D. This general method can be readily adapted for making corresponding 21-hydroxy derivatives from other compounds of the invention.
In another embodiment, Epo K, a P450 epoxidase that performs the epoxidation reaction in host cells that naturally produce epothilones or another epoxylase may be used to make 12,13-epoxy versions of the compounds of the present invention. A general method for using EpoK for epoxidation is described by Example 5 of PCT publication WO 00/31247 which is incorporated herein by reference. Example 27 describes in greater detail the epoxidation of 10,11-dehydroepothilone D to 10,11-dehydroepothilone B, the general method which can be readily adapted for making corresponding 12,13-epoxide derivatives from other compounds of the invention.
Alternatively, the epoxidation reaction can occur by contacting an epothilone compound containing a double bond at a position that corresponds to the bond between carbon-12 and carbon 13 to a culture of cells that expresses a functional Epo K. Such cells include the myxobacterium Sorangium cellulosum. In particularly preferred embodiments, the Sorangium cellulosum expresses Epo K but does not contain a functional epothilone polyketide synthase (xe2x80x9cPKSxe2x80x9d) gene. Such strains may be made by mutagenesis where one or more mutations in the epothilone PKS gene render it inoperative. Such mutants can occur naturally (which may be found by screening) or can be directed using either mutagens such as chemicals or irradation or by genetic manipulation. A particularly effective strategy for making strains with an inoperative epothilone PKS is homologous recombination as described by PCT publication WO 00/31247.
Formulation
A composition of the present invention generally comprises an inventive compound and a pharmaceutically acceptable carrier. The inventive compound may be free form or where appropriate as pharmaceutically acceptable derivatives such as prodrugs, and salts and esters of the inventive compound.
The composition may be in any suitable form such as solid, semisolid, or liquid form. See Pharmaceutical Dosage Forms and Drug Delivery Systems, 5th edition, Lippicott Williams and Wilkins (1991) which is incorporated herein by reference. In general, the pharmaceutical preparation will contain one or more of the compounds of the invention as an active ingredient in admixture with an organic or inorganic carrier or excipient suitable for external, enteral, or parenteral application. The active ingredient may be compounded, for example, with the usual non-toxic, pharmaceutically acceptable carriers for tablets, pellets, capsules, suppositories, pessaries, solutions, emulsions, suspensions, and any other form suitable for use. The carriers that can be used include water, glucose, lactose, gum acacia, gelatin, mannitol, starch paste, magnesium trisilicate, talc, corn starch, keratin, colloidal silica, potato starch, urea, and other carriers suitable for use in manufacturing preparations, in solid, semi-solid, or liquified form. In addition, auxiliary stabilizing, thickening, and coloring agents and perfumes may be used.
In one embodiment, the compositions containing an inventive compound are Cremophor(copyright)-free. Cremophor(copyright) (BASF Aktiengesellschaft) is a polyethoxylated castor oil which is typically used as a surfactant in formulating low soluble drugs. However, because Cremophor(copyright) can case allergic reactions in a subject, compositions that minimize or eliminate Cremophor(copyright) are preferred. Formulations of epothilone A or B that eliminate Cremophor(copyright) are described for example by PCT Publication WO 99/39694 which is incorporated herein by reference and may be adapted for use with the inventive compounds.
Where applicable, the inventive compounds may be formulated as microcapsules and nanoparticles. General protocols are described for example, by Microcapsules and Nanoparticles in Medicine and Pharmacy by Max Donbrow, ed., CRC Press (1992) and by U.S. Pat. Nos. 5,510,118; 5,534,270; and 5,662,883 which are all incorporated herein by reference. By increasing the ratio of surface area to volume, these formulations allow for the oral delivery of compounds that would not otherwise be amenable to oral delivery.
The inventive compounds may also be formulated using other methods that have been previously used for low solubility drugs. For example, the compounds may form emulsions with vitamin E or a PEGylated derivative thereof as described by WO 98/30205 and 00/71163which are incorporated herein by reference. Typically, the inventive compound is dissolved in an aqueous solution containing ethanol (preferably less than 1% w/v). Vitamin E or a PEGylated-vitamin E is added. The ethanol is then removed to form a pre-emulsion that can be formulated for intravenous or oral routes of administration. Another strategy involves encapsulating the inventive compounds in liposomes. Methods for forming liposomes as drug delivery vehicles are well known in the art. Suitable protocols include those described by U.S. Pat. Nos. 5,683,715; 5,415,869, and 5,424,073 which are incorporated herein by reference relating to another relatively low solubility cancer drug taxol and by PCT Publication WO 01/10412 which is incorporated herein by reference relating to epothilone B. Of the various lipids that may be used, particularly preferred lipids for making epothilone-encapsulated liposomes include phosphatidylcholine and polyethyleneglycol-derivitized distearyl phosphatidylethanolamine. Example 28 provides an illustrative protocol for making liposomes containing 10,11-dehydroepothilone D, the general method which can be readily adapted to make liposomes containing other compounds of the present invention.
Yet another method involves formulating the inventive compounds using polymers such as polymers such as biopolymers or biocompatible (synthetic or naturally occurring) polymers. Biocompatible polymers can be categorized as biodegradable and non-biodegradable. Biodegradable polymers degrade in vivo as a function of chemical composition, method of manufacture, and implant structure. Illustrative examples of synthetic polymers include polyanhydrides, polyhydroxyacids such as polylactic acid, polyglycolic acids and copolymers thereof, polyesters polyamides polyorthoesters and some polyphosphazenes. Illustrative examples of naturally occurring polymers include proteins and polysaccharides such as collagen, hyaluronic acid, albumin, and gelatin.
Another method involves conjugating the compounds of the present invention to a polymer that enhances aqueous solubility. Examples of suitable polymers include polyethylene glycol, poly-(d-glutamic acid), poly-(1-glutamic acid), poly-(1-glutamic acid), poly-(d-aspartic acid), poly-(1-aspartic acid), poly-(1-aspartic acid) and copolymers thereof. Polyglutamic acids having molecular weights between about 5,000 to about 100,000 are preferred, with molecular weights between about 20,000 and 80,000 being more preferred and with molecular weights between about 30,000 and 60,000 being most preferred. The polymer is conjugated via an ester linkage to one or more hydroxyls of an inventive epothilone using a protocol as essentially described by U.S. Pat. No. 5,977,163 which is incorporated herein by reference, and by Example 29. Preferred conjugation sites include the hydroxyl off carbon-21 in the case of 21-hydroxy-derivatives of the present invention. Other conjugation sites include the hydroxyl off carbon 3 and the hydroxyl off carbon 7.
In another method, the inventive compounds are conjugated to a monoclonal antibody. This strategy allows the targeting of the inventive compounds to specific targets. General protocols for the design and use of conjugated antibodies are described in Monoclonal Antibody-Based Therapy of Cancer by Michael L. Grossbard, ed. (1998) which is incorporated herein by reference.
The amount of active ingredient that may be combined with the carrier materials to produce a single dosage form will vary depending upon the subject treated and the particular mode of administration. For example, a formulation for intravenous use comprises an amount of the inventive compound ranging from about 1 mg/mL to about 25 mg/mL, preferably from about 5 mg/mL to 15 mg/mL, and more preferably about 10 mg/mL. Intravenous formulations are typically diluted between about 2 fold and about 30 fold with normal saline or 5% dextrose solution prior to use.
Methods to Treat Cancer
In one aspect of the present invention, the inventive compounds are used to treat cancer. In one embodiment, the compounds of the present invention are used to treat cancers of the head and neck which include tumors of the head, neck, nasal cavity, paranasal sinuses, nasopharynx, oral cavity, oropharynx, larynx, hypopharynx, salivary glands, and paragangliomas. In another embodiment, the compounds of the present invention are used to treat cancers of the liver and biliary tree, particularly hepatocellular carcinoma. In another embodiment, the compounds of the present invention are used to treat intestinal cancers, particularly colorectal cancer. In another embodiment, the compounds of the present invention are used to treat ovarian cancer. In another embodiment, the compounds of the present invention are used to treat small cell and non-small cell lung cancer. In another embodiment, the compounds of the present invention are used to treat breast cancer. In another embodiment, the compounds of the present invention are used to treat sarcomas which includes fibrosarcoma, malignant fibrous histiocytoma, embryonal rhabdomysocarcoma, leiomysosarcoma, neurofibrosarcoma, osteosarcoma, synovial sarcoma, liposarcoma, and alveolar soft part sarcoma. In another embodiment, the compounds of the present invention are used to treat neoplasms of the central nervous systems, particularly brain cancer. In another embodiment, the compounds of the present invention are used to treat lymphomas which include Hodgkin""s lymphoma, lymphoplasmacytoid lymphoma, follicular lymphoma, mucosa-associated lymphoid tissue lymphoma, mantle cell lymphoma, B-lineage large cell lymphoma, Burkitt""s lymphoma, and T-cell anaplastic large cell lymphoma.
The method comprises administering a therapeutically effective amount of an inventive compound to a subject suffering from cancer. The method may be repeated as necessary either to contain (i.e. prevent further growth) or to eliminate the cancer. Clinically, practice of the method will result in a reduction in the size or number of the cancerous growth and/or a reduction in associated symptoms (where applicable). Pathologically, practice of the method will produce at least one of the following: inhibition of cancer cell proliferation, reduction in the size of the cancer or tumor, prevention of further metastasis, and inhibition of tumor angiogenesis.
The compounds and compositions of the present invention can be used in combination therapies. In other words, the inventive compounds and compositions can be administered concurrently with, prior to, or subsequent to one or more other desired therapeutic or medical procedures. The particular combination of therapies and procedures in the combination regimen will take into account compatibility of the therapies and/or procedures and the desired therapeutic effect to be achieved.
In one embodiment, the compounds and compositions of the present invention are used in combination with another anti-cancer agent or procedure. Illustrative examples of other anti-cancer agents include but are not limited to: (i) alkylating drugs such as mechlorethamine, chlorambucil, Cyclophosphamide, Melphalan, Ifosfamide; (ii) antimetabolites such as methotrexate; (iii) microtubule stabilizing agents such as vinblastin, paclitaxel, docetaxel, and discodermolide; (iv) angiogenesis inhibitors; (v) and cytotoxic antibiotics such as doxorubicon (adriamycin), bleomycin, and mitomycin. Illustrative examples of other anti-cancer procedures include: (i) surgery; (ii) radiotherapy; and (iii) photodynamic therapy.
In another embodiment, the compounds and compositions of the present invention are used in combination with an agent or procedure to mitigate potential side effects from the inventive compound or composition such as diarrhea, nausea and vomiting. Diarrhea may be treated with antidiarrheal agents such as opioids (e.g. codeine, diphenoxylate, difenoxin, and loeramide), bismuth subsalicylate, and octreotide. Nausea and vomiting may be treated with antiemetic agents such as dexamethasone, metoclopramide, diphenyhydramine, lorazepam, ondansetron, prochlorperazine, thiethylperazine, and dronabinol. For those compositions that includes polyethoxylated castor oil such as Cremophorg(copyright), pretreatment with corticosteroids such as dexamethasone and methylprednisolone and/or H1 antagonists such as diphenylhydramine HCl and/or H2 antagonists may be used to mitigate anaphylaxis. Illustrative formulations for intravenous use and pretreatment regiments are described by Examples 30 and 31 respectively.
Methods of Treating of Non-cancer, Cellular Hyperproliferative Disorders
In another aspect of the present invention, the inventive compounds are used to treat non-cancer disorders that are characterized by cellular hyperproliferation. In one embodiment, the compounds of the present invention are used to treat psoriasis, a condition characterized by the cellular hyperproliferation of keratinocytes which builds up on the skin to form elevated, scaly lesions. The method comprises administering a therapeutically effective amount of an inventive compound to a subject suffering from psoriasis. The method may be repeated as necessary either to decrease the number or severity of lesions or to eliminate the lesions. Clinically, practice of the method will result in a reduction in the size or number of skin lesions, diminution of cutaneous symptoms (pain, burning and bleeding of the affected skin) and/or a reduction in associated symptoms (e.g., joint redness, heat, swelling, diarrhea. abdominal pain, Pathologically, practice of the method will result in at least one of the following: inhibition of keratinocyte proliferation, reduction of skin inflammation (for example, by impacting on: attraction and growth factors, antigen presentation, production of reactive oxygen species and matrix metalloproteinases), and inhibition of dermal angiogenesis.
In another embodiment, the compounds of the present invention are used to treat multiple sclerosis, a condition characterized by progressive demyelination in the brain. Although the exact mechanisms involved in the loss of myelin are not understood, there is an increase in astrocyte proliferation and accumulation in the areas of myelin destruction. At these sites, there is macrophage-like activity and increased protease activity which is at least partially responsible for degradation of the myelin sheath. The method comprises administering a therapeutically effective amount of an inventive compound to a subject suffering from multiple sclerosis. The method may be repeated as necessary to inhibit astrocyte proliferation and/or lessen the severity of the loss of motor function and/or prevent or attenuate chronic progression of the disease. Clinically, practice of the method will result in in improvement in visual symptoms (visual loss, diplopia), gait disorders (weakness, axial instability, sensory loss, spasticity, hyperreflexia, loss of dexterity), upper extremity dysfunction (weakness, spasticity, sensory loss), bladder dysfunction (urgency, incontinence, hesitancy, incomplete emptying), depression, emotional lability, and cognitive impairment. Pathologically, practice of the method will result in the reduction of one or more of the following, such as myelin loss, breakdown of the blood-brain barrier, perivascular infiltration of mononuclear cells, immunologic abnormalities, gliotic scar formation and astrocyte proliferation, metalloproteinase production, and impaired conduction velocity.
In another embodiment, the compounds of the present invention are used to treat rheumatoid arthritis, a multisystem chronic, relapsing, inflammatory disease that sometimes leads to destruction and ankyiosis of affected joints. Rheumatoid arthritis is characterized by a marked thickening of the synovial membrane which forms villous projections that extend into the joint space, multilayering of the synoviocyte lining (synoviocyte proliferation), infiltration of the synovial membrane with white blood cells (macrophages, lymphocytes, plasma cells, and lymphoid follicles; called an xe2x80x9cinflammatory synovitisxe2x80x9d), and deposition of fibrin with cellular necrosis within the synovium. The tissue formed as a result of this process is called pannus and, eventually the pannus grows to fill the joint space. The pannus develops an extensive network of new blood vessels through the process of angiogenesis that is essential to the evolution of the synovitis. Release of digestive enzymes (matrix metalloproteinases (e.g., collagenase, stromelysin)) and other mediators of the inflammatory process (e.g., hydrogen peroxide, superoxides, lysosomal enzymes, and products of arachadonic acid metabolism) from the cells of the pannus tissue leads to the progressive destruction of the cartilage tissue. The pannus invades the articular cartilage leading to erosions and fragmentation of the cartilage tissue. Eventually there is erosion of the subchondral bone with fibrous ankylosis and ultimately bony ankylosis, of the involved joint.
The method comprises administering a therapeutically effective amount of an inventive compound to a subject suffering from rheumatoid arthritis. The method may be repeated as necessary to accomplish to inhibit synoviocyte proliferation and/or lessen the severity of the loss of movement of the affected joints and/or prevent or attenuate chronic progression of the disease. Clinically, practice of the present invention will result in one or more of the following: (i) decrease in the severity of symptoms (pain, swelling and tenderness of affected joints; morning stiffness, weakness, fatigue, anorexia, weight loss); (ii) decrease in the severity of clinical signs of the disease (thickening of the joint capsule, synovial hypertrophy, joint effusion, soft tissue contractures, decreased range of motion, ankylosis and fixed joint deformity); (iii) decrease in the extra-articular manifestations of the disease (rheumatic nodules, vasculitis, pulmonary nodules, interstitial fibrosis, pericarditis, episcleritis, iritis, Felty""s syndrome, osteoporosis); (iv) increase in the frequency and duration of disease remission/symptom-free periods; (v) prevention of fixed impairment and disability; and/or (vi) prevention/attenuation of chronic progression of the disease. Pathologically, practice of the present invention will produce at least one of the following: (i) decrease in the inflammatory response; (ii) disruption of the activity of inflammatory cytokines (such as IL-I, TNFa, FGF, VEGF); (iii) inhibition of synoviocyte proliferation; (iv) inhibition of matrix metalloproteinase activity, and/or (v) inhibition of angiogenesis.
In another embodiment, the compounds of the present invention are used to threat atherosclerosis and/or restenosis, particularly in patients whose blockages may be treated with an endovascular stent. Atheroschlerosis is a chronic vascular injury in which some of the normal vascular smooth muscle cells (xe2x80x9cVSMCxe2x80x9d) in the artery wall, which ordinarily control vascular tone regulating blood flow, change their nature and develop xe2x80x9ccancer-likexe2x80x9d behavior. These VSMC become abnormally proliferative, secreting substances (growth factors, tissue-degradation enzymes and other proteins) which enable them to invade and spread into the inner vessel lining, blocking blood flow and making that vessel abnormally susceptible to being completely blocked by local blood clotting. Restenosis, the recurrence of stenosis or artery stricture after corrective procedures, is an accelerated form of atherosclerosis.
The method comprises coating a therapeutically effective amount of an inventive compound on a stent and delivering the stent to the diseased artery in a subject suffering from atherosclerosis. Methods for coating a stent with a compound are described for example by U.S. Pat. Nos. 6,156,373 and 6,120, 847. Clinically, practice of the present invention will result in one or more of the following: (i) increased arterial blood flow; (ii) decrease in the severity of clinical signs of the disease; (iii) decrease in the rate of restenosis; or (iv) prevention/attenuation of the chronic progression of atherosclerosis. Pathologically, practice of the present invention will produce at least one of the following at the site of stent implanataion: (i) decrease in the inflammatory response, (ii) inhibition of VSMC secretion of matrix metalloproteinases; (iii) inhibition of smooth muscle cell accumulation; and (iv) inhibition of VSMC phenotypic dedifferentiation.
Dosage Levels
In one embodiment, dosage levels that are administered to a subject suffering from cancer or a non-cancer disorder characterized by cellular proliferation are of the order from about 1 mg/m2 to about 200 mg/m2 which may be administered as a bolus (in any suitable route of administration) or a continuous infusion (e.g. 1 hour, 3 hours, 6 hours, 24 hours, 48 hours or 72 hours) every week, every two weeks, or every three weeks as needed. It will be understood, however, that the specific dose level for any particular patient depends on a variety of factors. These factors include the activity of the specific compound employed; the age, body weight, general health, sex, and diet of the subject; the time and route of administration and the rate of excretion of the drug; whether a drug combination is employed in the treatment; and the severity of the condition being treated.
In another embodiment, the dosage levels are from about 10 mg/m2 to about 150 mg/m2, preferably from about 10 to about 75 mg/m2 and more preferably from about 15 mg/m2 to about 50 mg/m2 once every three weeks as needed and as tolerated. In another embodiment, the dosage levels are from about 1 mg to about 150 mg/m2, preferably from about 10 mg/m2 to about 75 mg/m2 and more preferably from about 25 mg/m2 to about 50 mg/m2 once every two weeks as needed and as tolerated. In another embodiment, the dosage levels are from about 1 mg/m2 to about 100 mg/m2, preferably from about 5 mg/m2 to about 50 mg/m2 and more preferably from about 10 mg/m2 to about 25 mg/m2 once every week as needed and as tolerated. In another embodiment, the dosage levels are from about 0.1 to about 25 mg/m2, preferably from about 0.5 to about 15 mg/m2 and more preferably from about 1 mg/m2 to about 10 mg/m2 once daily as needed and tolerated.
A detailed description of the invention having been provided above, the following examples are given for the purpose of illustrating the present invention and shall not be construed as being a limitation on the scope of the invention or claims.